Inefficient assembly and intracellular accumulation of antibodies with mutations in V(H) CDR2

Tammy Martin, Gregory D. Wiens, Marvin Rittenberg

Research output: Contribution to journalArticle

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Abstract

We previously described secretion defects in four mutants of the murine anti-phosphocholine Ab, T15. The mutant heavy (H) chains had amino acid replacements in the V(H) complementarity-determining region 2 (HCDR2) and were expressed at normal intracellular levels. Here, the intracellular fate of the secretion-defective mutant heavy chains was investigated. Metabolic labeling demonstrated that the T15 wild-type Ab was secreted within a 4-h chase. In contrast, the mutant H chains accumulated with intracellular t(1/2) values ranging from 10 to 24 h. The mutant H chains were associated with increased levels of the molecular chaperones BiP and GRP94, and remained endoglycosidase H sensitive, suggesting retention in the endoplasmic reticulum. Assembly of the mutant H chains with T15 light (L) chain was arrested at the H2 and H2L intermediate stages of the T15 wild-type pathway (H2 → H2L → H2L2). Even though some assembly with L chain occurred, it was not as a secretion-competent H2L2 Ig moiety. The T15 L chains coexpressed with mutant H chains were degraded efficiently except for a minor L chain population with a long t(1/2) that was apparently protected at the H2L stage. To our knowledge, this is the first study demonstrating that intracellular half-lives of Ig H and L chains can be influenced by somatic mutations in HCDR2.

Original languageEnglish (US)
Pages (from-to)5963-5970
Number of pages8
JournalJournal of Immunology
Volume160
Issue number12
StatePublished - Jun 15 1998

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Complementarity Determining Regions
Mutation
Phosphorylcholine
Antibodies
Glycoside Hydrolases
Endoplasmic Reticulum
Light
Amino Acids
Population
glucose-regulated proteins
molecular chaperone GRP78

ASJC Scopus subject areas

  • Immunology

Cite this

Inefficient assembly and intracellular accumulation of antibodies with mutations in V(H) CDR2. / Martin, Tammy; Wiens, Gregory D.; Rittenberg, Marvin.

In: Journal of Immunology, Vol. 160, No. 12, 15.06.1998, p. 5963-5970.

Research output: Contribution to journalArticle

Martin, Tammy ; Wiens, Gregory D. ; Rittenberg, Marvin. / Inefficient assembly and intracellular accumulation of antibodies with mutations in V(H) CDR2. In: Journal of Immunology. 1998 ; Vol. 160, No. 12. pp. 5963-5970.
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